Study On Therapeutic Effect Of Zhizhu Decoction On Slow Transit Constipation And Explanation Of Its Mechanism

Objective:1.To evaluate the efficacy and safety of Zhizhu prescription and prucalopride in the treatment of chronic constipation,the systematic reviews of randomized controlled trails(RCTs)and clinical controlled trials(CCTs)of these drugs are performed.2.Selecting Zhishi and Baizhu as the research objects and prucalopride as control group,to observe the interventional effects of the different proportions of couple of Zhishi and Baizhu on the intestinal movements of the in the model rats with slow transit constipation,and try to find the best ones.To observe the effect on the signaling pathways of 5-HT3、5-HT4 and neurotransmitters in the gut,evaluate the safety,and investigate initially the effects and plausible mechanism of Zhizhu prescription on slow transit constipation.Methods:1.Systematic reviewsSearch through the RCTs and CCTs of the treatment of the chronic constipation with Zhizhu prescription and prucalopride in the PubMed database,Embase medicine database,OVID database,SinoMed,VIP,Wanfang Chinese Medical Association Periodical and CNKI.To provide a basis of the evidence-based medicine,these RCTs and CCTs are through systematic literature reviews and meta-analysises.2.Experimental study(1)Experiment OneThe 80 healthy SD rats are randomly assigned to two groups including healthy group 10 rats and slow transit constipation model group 70 rats made by "Rhein suspension gavage method".And the 70 rats are randomly assigned to seven groups including model group 10 rats and six treatment groups with different kinds of compatibility proportions of Zhizhu prescriptions.After the modeling,the treatment groups have perfused with 6 kinds of compatibility proportions of Zhizhu prescriptions respectively for 14 days,and healthy group and model group with natural drink and food.Then the intestinal transmission function is examined,and the results of the collected data are compared.Try to find the best proportion of couple of Zhishi and Baizhu from the different prescriptions.(2)Experiment TwoThe 40 healthy SD rats are randomly assigned to four groups including healthy group,model group,treatment group and control group.The slow transit constipation animal models are made on all of rats by "Rhein suspension gavage method" except healthy group.After the modeling,the treatment group has perfused with Zhizhu prescription for 14 days,and the control group has perfused with prucalopride suspension for 14 days,and healthy group and model group with natural drink and food.Then the intestinal transmission function and fecal water content are examined.After administration,liver and kidney function is measured.The tissues of stomach,heart,liver,kidney and proximal colon are collected to make a pathological observation by using light microscope.Liver and kidney are weighed to work out the liver coefficient and kidney coefficient.(3)Experiment Three5-HT,5-HT3 receptors,5-HT4 receptors,SERT,ChAT,VIP and SP are positioned in histology by immunohistochemical method.The level expression of mRNA of 5-HT3 receptors,5-HT4 receptors,SERT,ChAT,VIP and SP in colon tissue are detected by RealTime-PCR.Results:1.Zhizhu prescription for the treatment of chronic constipation:a systematic reviewThrough screening,10 literatures(all are in Chinese)involving 1382 adults are meet the standards finally.One of them is CCT,and the rest are RCTs.8 studies report cure rates,10 studies report overall efficiency rates,2 studies report the changes in colonic transit function test,3 studies report the scores of constipation,5 studies report recurrence rates,3 studies report adverse events.Meta-analysis shows that the treatment groups are better than control groups in cure rates,overall efficiency rates and recurrence rates.The results of colonic transit function test report both the treatment groups and the control groups have no significant difference or the treatment groups are better than the control groups.The scores of constipation report both of the treatment groups and the control groups can improve constipation,but which are the better ones,the uniform conclusion can’t to be found.The adverse reactions are mainly manifested as diarrhea,abdominal pain,et al.But they graduated away after stopping the treatment.2.Prucalopride for the treatment of chronic constipation:a systematic reviewThrough screening,5 RCTs(1 of them is in Chinese,and 4 of them are in English)involving 2587 adults are meet the standards finally.5 studies report the proportion of patients with≥3 SCBMs/week,3 studies report overall PAC-SYM symptoms score.3 studies report the proportion of patients with improvement≥1 PAC-QOL satisfaction score from baseline.5 studies report adverse events.Meta-analysis shows that the proportion of patients with≥3 SCBMs/week,overall PAC-SYM symptoms score and the proportion of patients with improvement≥1 PAC-QOL satisfaction score from baseline were significantly better with prucalopride 2mg/d and 4mg/d compared with placebo.There are no differences between the two groups.The adverse reactions are mainly manifested as nausea,diarrhea and headache.The proportion of patients dropped out of the study was significantly lower with prucalopride 2mg/d compared with prucalopride 4mg/d.3.Experimental studyAfter 110 days,slow transit constipation rat models are made successfully.The best optimal dosage is 30g Zhishi and 70g Baizhu.The ink propelling rates is the highest in this case.Both of the fecal water contents of the treatment group and the control group are slightly increased compared with model group.But there are no differences between the two groups(P>0.05).The ink propelling rates of treatment group and control group are significantly increased compared with model group(P<0.01),but there are no differences between the two groups(P>0.05).The results report that Zhizhu prescription and Prucalopride can’t increase the fecal water content.They improve constipation through promoting colonic transit function.No treatment-related abnormal changes as liver and kidney function,liver coefficient and kidney coefficient are observed and mo toxic pathological changes of pathomorphology occurred in the tissues of stomach,heart,liver and kidney.The results report that Zhizhu prescription and Prucalopride are safe and effective in the treatment of slow transit constipation.The number of nerve cells in the bowel layers of the proximal colon decrease and they show vacuolation degeneration,which are detectd by pathological test.After 14 days of treatment and medication,the degree of pathological changes is reduced.Results show that both of the Zhizhu prescription and prucalopride have some therapeutic effects on the model rats.Immunohistochemical analysis show that the 5-HT expression levels have elevated and the expression levels of SERT、5-HT3R,5-HT4R,ChAT,VIP and SP have dropped in the colon tissue of model rats.But compare with model group,the 5-HT expression levels have dropped and the expression levels of SERT、5-HT3R,5-HT4R,ChAT,VIP and SP have elevated in the colon tissue of the treatment group and control group.The effect of Zhizhu prescription is little worse in prucalopride.The results of Real Time-PCR assay parallel the results of mmunohistochemistry.Conclusion:1.According to the meta-analysis of these ten trails,Zhizhu prescription was safe and efective in the treatment of chronic constipation,which can’t be strongly proved at present for lack of studies with high quality.2.According to the meta-analysis of these 5 trails,prucalopride 2mg/d was safe and effective in the treatment of chronic constipation,which can’t be strongly proved at present for lack of more studies with high quality.3、The optimal proportion of Zhizhu prescription in the treatment of slow transit constipation is almost 1:2.However for the limited conditions of experimental design,there are to be lack of animal experiments about Zhizhu prescription that proportion is 1:1,single Zhishi prescription,single Baizhu prescription and so on.Additionally,when the Zhizhu prescription fixes the proportion as 1:2,the experimental results show a distinct dose-and-effect relationship.The study result shows that the best dose of Zhizhu prescription is 30g Zhishi and 70g Baizhu.But further research is needed to see whether the dose would have its maximum impact.The results report that Zhizhu prescription and Prucalopride can’t increase the fecal water content.They improve constipation through promoting colonic transit function.The results report that Zhizhu prescription and Prucalopride are safe and effective in the treatment of slow transit constipation.4、The mechanism of the treatment on slow transit constipation by Zhizhu prescription therapy may be related to the repair of signaling the abnormalities in the signaling pathways of 5-HT3R、5-HT4R and neurotransmitters in the gut.